Key Product Details

Species Reactivity

Validated:

Multi-Species

Cited:

Human, Mouse, Rat, Transgenic Mouse

Applications

Validated:

Immunohistochemistry, Intracellular Staining by Flow Cytometry, Immunocytochemistry

Cited:

Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot, Flow Cytometry, Immunocytochemistry, Immunoprecipitation, Chromatin Immunoprecipitation (ChIP), Co-Immunoprecipitation, ELISA Capture, Proximity Ligation Assay, Functional Assay, Microarray, Westen Blot

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG3 Clone # 10HA
View all PAR/pADPr Primary Antibodies »

Product Specifications

Immunogen

Purified ADP-ribose polymers between 2 and 50 units long

Specificity

The antibody is specific for PAR polymers 2 to 50 units long, but does not recognize structurally related RNA, DNA, ADP-ribose monomers, NAD, or other nucleic acid monomers.  Detects Poly(ADP-ribose) Polymer in Direct ELISA.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG3

Scientific Data Images for PAR/pADPr Antibody

Detection of PAR in Jurkat cells by Flow Cytometry.

Jurkat human acute T cell leukemia cell line treated with 1 mM H2O2 for 5 minutes (filled histogram) or resting (open histogram) were stained with Mouse Anti-PAR Monoclonal Antibody (Catalog # 4335-MC-100) followed by Goat anti-Mouse IgG PE-conjugated Secondary Antibody (F0102B). To facilitate intracellular staining, cells were fixed and permeabilized using FlowX FoxP3/Transcription Factor Fixation & Perm Buffer Kit (FC012). Staining was performed using our Staining Intracellular Molecules protocol.

Detection of PAR/pADPr in Human Kidney.

PAR/pADPr was detected in immersion fixed paraffin-embedded sections of human kidney using Mouse Anti-PAR/pADPr Monoclonal Antibody (Catalog # 4335-MC-100) at 1.7 µg/ml for 1 hour at room temperature followed by incubation with the Anti-Mouse IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC001). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using VisUCyte Antigen Retrieval Reagent-Basic (Catalog # VCTS021). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to the nucleus. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

Detection of PAR/pADPr in 786‑O Human Cell Line.

PAR/pADPr was detected in immersion fixed 786‑O human renal cell adenocarcinoma cell line untreated (negative) or treated (positive) with 2mM hydrogen peroxide for 5 minutes using Mouse Anti-PAR/pADPr Monoclonal Antibody (Catalog # 4335-MC-100) at 3 µg/ml for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to the nucleus upon stimulation with hydrogen peroxide. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Detection of PAR/pADPr by Western Blot

Detection of PAR/pADPr by Western Blot

PARP1 promotes PARP2 recruitment at DNA damage sites and niraparib enhances PARP2 foci independent of PARP1. (A) Representative images of laser-induced GFP-PARP2 and mRFP-XRCC1 foci in WT, Parp1 KO, Parp2 KO, and Parp1/2 DKO iMEF cells. The yellow arrowheads point to the area of micro-irradiation. (B) The relative intensity kinetics and (C) The normalized kinetics (plotted as the percentage of the maximal relative intensity at 1 min) of GFP-PARP2 at DNA damage sites in WT, Parp1 KO, Parp2 KO and Parp1/2 DKO iMEF cells. (D, E) The relative intensity of (B) GFP-PARP2 and (C) RFP-XRCC1 at 1 min post-microirradiation. Two-tailed unpaired Student's t-test were used to calculate the p-values. (F) Soluble and chromatin fraction of WT or PARP1 KO RPE-1 cells treated with MMS (0.1 mg/ml, 1 h) and niraparib (1 μM, 1 h). (G) The niraparib sensitivity of WT, PARP1 KO, PARP2 KO, and PARP1/2 DKO RPE-1 cells. The dots and error bars represent means and SEM, respectively, from one representative experiment out of three consistent biological repeats with triplicate samples (for sensitivity) and at least 8–10 cells (for live cell imaging) per experiment. P value of IC50 was calculated using the extra sum-of-square F test. ns, P > 0.05; **P < 0.01; ***P < 0.001. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/35349716), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of PAR/pADPr by Western Blot

Detection of PAR/pADPr by Western Blot

180055 blocks the catalytic activity of PARP1.A–C T47D (A), MDA-MB-231 (B), and MOLT4 cells (C) were pre-treated with Rucaparib or 180055 (1 μM) for 24 h and then challenged with H2O2 (2 mM) for 1 h. NAD+ levels were then determined; values represent mean ± SEM (n = 3 biological independent samples). Statistical significance was calculated with one-way ANOVA comparing the H2O2-treated DMSO group to the Rucaparib and 180055 groups. ***p < 0.001, ****p < 0.0001. D–F immunoblot analysis of PARylation levels in T47D (D), MDA-MB-231 (E), and MOLT4 cells (F) treated with either Rucaparib or 180055. Cells were pre-treated with a PARG inhibitor (PDD 00017273, 2 μM) for 1 h and then treated with Rucaparib or 180055 (1 μM) for 1 h. Cells were then challenged with H2O2 (2 mM) for 5 min, and whole cell lysates were analyzed using immunoblot assays with the indicated antibodies. Image collected and cropped by CiteAb from the following open publication (https://www.nature.com/articles/s41419-024-07277-2), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of PAR/pADPr by Western Blot

Detection of PAR/pADPr by Western Blot

180055 blocks the catalytic activity of PARP1.A–C T47D (A), MDA-MB-231 (B), and MOLT4 cells (C) were pre-treated with Rucaparib or 180055 (1 μM) for 24 h and then challenged with H2O2 (2 mM) for 1 h. NAD+ levels were then determined; values represent mean ± SEM (n = 3 biological independent samples). Statistical significance was calculated with one-way ANOVA comparing the H2O2-treated DMSO group to the Rucaparib and 180055 groups. ***p < 0.001, ****p < 0.0001. D–F immunoblot analysis of PARylation levels in T47D (D), MDA-MB-231 (E), and MOLT4 cells (F) treated with either Rucaparib or 180055. Cells were pre-treated with a PARG inhibitor (PDD 00017273, 2 μM) for 1 h and then treated with Rucaparib or 180055 (1 μM) for 1 h. Cells were then challenged with H2O2 (2 mM) for 5 min, and whole cell lysates were analyzed using immunoblot assays with the indicated antibodies. Image collected and cropped by CiteAb from the following open publication (https://www.nature.com/articles/s41419-024-07277-2), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Mouse PAR/pADPr by Western Blot

Detection of Mouse PAR/pADPr by Western Blot

Regulation of NAD+-consuming enzymes PARP1 and PAR-AIF by Ginsenoside Rb1 in OGD/R-injured ASs. (A,B) Western blot analysis and quantification of PARP1 protein levels. (C,D) Western blot analysis and quantification of PAR. The band intensity of each protein was plotted after normalizing to the beta -tubulin signal of the same lane. Data are shown with mean ± SD, n = 3. ##p < 0.01 vs. Control group, * p < 0.05, ** p < 0.01 vs. OGD/R group based on one-way ANOVA with Fisher’s LSD test. (E,F) Immunofluorescence shows PAR expression in ASs. PAR (green—FITC) and nuclei stained by DAPI are shown, as well as the merged images. The bar graphs (mean ± SD, n = 5) represent the quantitative results of fluorescence intensity in 3 different fields for each group. (G,H) Immunofluorescence shows the distribution of AIF to the nucleus. AIF (green—FITC) and nucleus stained by DAPI are shown, as well as the merged images. Co-immunofluorescence analysis revealed that the nuclear translocation of AIF in OGD/R-injured ASs was blocked by Rb1 treatment. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/38003249), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of PAR/pADPr by Western Blot

Detection of PAR/pADPr by Western Blot

180055 blocks the catalytic activity of PARP1.A–C T47D (A), MDA-MB-231 (B), and MOLT4 cells (C) were pre-treated with Rucaparib or 180055 (1 μM) for 24 h and then challenged with H2O2 (2 mM) for 1 h. NAD+ levels were then determined; values represent mean ± SEM (n = 3 biological independent samples). Statistical significance was calculated with one-way ANOVA comparing the H2O2-treated DMSO group to the Rucaparib and 180055 groups. ***p < 0.001, ****p < 0.0001. D–F immunoblot analysis of PARylation levels in T47D (D), MDA-MB-231 (E), and MOLT4 cells (F) treated with either Rucaparib or 180055. Cells were pre-treated with a PARG inhibitor (PDD 00017273, 2 μM) for 1 h and then treated with Rucaparib or 180055 (1 μM) for 1 h. Cells were then challenged with H2O2 (2 mM) for 5 min, and whole cell lysates were analyzed using immunoblot assays with the indicated antibodies. Image collected and cropped by CiteAb from the following open publication (https://www.nature.com/articles/s41419-024-07277-2), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for PAR/pADPr Antibody

Application
Recommended Usage

Immunocytochemistry

3-25 µg/mL
Sample: Immersion fixed 786-O human renal cell adenocarcinoma cell line

Immunohistochemistry

1-15 µg/mL
Sample: Immersion fixed paraffin-embedded sections of human kidney

Intracellular Staining by Flow Cytometry

0.25 µg/106 cells
Sample: Jurkat human acute T cell leukemia cell line treated with 1 mM H2O2 for 5 minutes

Reviewed Applications

Read 3 reviews rated 4 using 4335-MC-100 in the following applications:

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Flow Cytometry

Formulation, Preparation, and Storage

Purification

Protein A or G purified from ascites

Formulation

Supplied as a 0.2 μm filtered solution in TBS. See Certificate of Analysis for details.

Shipping

The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C, as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after opening.
  • 6 months, -20 to -70 °C under sterile conditions after opening.

Background: PAR/pADPr

PARP [Poly(ADP-ribose) Polymerase], also known as ADPRT and PPOL, is a 118-kDa enzyme that uses NAD as a substrate to catalyze the covalent transfer of ADP-ribose to a variety of nuclear protein acceptors. ADP ribosyltransferase is required for cellular repair, and PARP expression is induced by single-strand breaks in DNA. PARP is proteolytically cleaved by Caspase-3 into two fragments of 89- and 24-kDa in one of the hallmark events of apoptosis.

Long Name

Poly [ADP-ribose] Polymer

Alternate Names

PAR

Additional PAR/pADPr Products

Product Documents for PAR/pADPr Antibody

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Note: Certificate of Analysis not available for kit components.

Product Specific Notices for PAR/pADPr Antibody

For research use only

Related Research Areas

Apoptosis

Citations for PAR/pADPr Antibody

Customer Reviews for PAR/pADPr Antibody (3)

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Customer Images

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  • PAR/pADPr Antibody
    Name: Anonymous
    Application: Western Blot
    Sample Tested: THP-1 human acute monocytic leukemia cell line
    Species: Human
    Verified Customer | Posted 03/15/2024
    SDS-PAGE 10% gel of THP-1 monocytes with/without 5 mins 2mM H2O2 treatment. 30ug of protein run. Anti-PAR/pADPr used at 1:2,000 diln in 5% BSA in TBS-T for 1h.
    PAR/pADPr Antibody 4335-MC-100
  • PAR/pADPr Antibody
    Name: Anonymous
    Application: Immunoprecipitation
    Sample Tested: THP-1 human acute monocytic leukemia cell line
    Species: Human
    Verified Customer | Posted 03/08/2024
    Antibody was used to perform immunoprecipitation (2ug) of THP-1 lysates before SDS-PAFE of samples and immunoblot was performed with anti-CTCF (1:5k diln). Parylated-CTCF found at 180kDa in righthand lane.
    PAR/pADPr Antibody 4335-MC-100
  • PAR/pADPr Antibody
    Name: Marina Rodriguez
    Application: Western Blot
    Sample Tested: Breast cancer cells
    Species: Human
    Verified Customer | Posted 02/02/2021
    8% polyacrylamide gel run until 55 kDa band. 1:750 primary antibody dilution; 1:10000 secondary antibody dilution. UT=untreated cells.
    PAR/pADPr Antibody 4335-MC-100

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Protocols

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FAQs for PAR/pADPr Antibody

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    • A: The PAR/pADPr Antibody (Catalog # 4335-MC-100) is supplied as a solution in TBS containing glycerol, at a concentration of 100 µg/100 µL.
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